CN106811423B - Germanium-rich yeast and preparation method and application thereof - Google Patents
Germanium-rich yeast and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses germanium-rich yeast and a preparation method and application thereof. Firstly, carrying out subculture and acclimatization on candida utilis in a germanium-containing culture medium to obtain acclimatized candida utilis; then, the domesticated candida utilis is cultured in a yeast fermentation culture medium to obtain germanium-rich yeast. The invention utilizes the germanium-rich capability of candida utilis and the capability of cheap raw materials, and finally obtains the germanium-rich yeast with high biological activity and low toxicity by taking molasses and urea as fermentation raw materials. By adopting the method, the germanium-rich yeast product with high purity and yield of organic germanium can be finally obtained, wherein the germanium content in each kilogram of the germanium-rich yeast product is 837.1-1206.4 mg, and the organic germanium content reaches 93.3% -98.6%. The method can convert inorganic germanium into organic germanium, expand the source of organic germanium, and promote the large-scale application of germanium in the fields of food, feed and the like.
Description
Technical Field
The invention belongs to the field of food and feed additives, and relates to germanium-rich yeast and a preparation method and application thereof.
Background
Germanium was found by Winkler, a german chemist, in 1886 as a trace element. In 1929, Dutoit found germanium in animals. Many enzymes in animals (e.g., guanadinase, cytochrome oxidase), brain cortex and gray matter components contain trace amounts of germanium. After the bacteriostatic activity of trialkyl germanium acetate is discovered in 1962, the biological effect of organic germanium attracts attention. Currently, some compounds of germanium have demonstrated various biological activities.
Biological function of germanium:
(1) aso participating in immune regulation is found through a mouse test that germanium can obviously improve the content of interferon in mouse serum. According to reports of Tangmeganew and the like, the plasma antibody level of chicks produced by feeding the breeding hens with the germanium-containing feed is doubled compared with that of a control group, and after the chicks are fed for a period of time, the thymus index, the bursa melting index and the spleen index are also obviously higher than those of the control group. Studies of Chengyi, historic literature, etc. have shown that germanium enhances the conversion of T lymphocytes in chicken blood.
(2) The bacteriostatic germanium has strong bacteriostatic action. Karrs and Sijpertein demonstrated that trialkyl acetyl germanium has antimicrobial activity, and the main active compound in trialkyl acetyl germanium for inhibiting fungi is ethyl, propyl germanium compound, the longer its chain, the stronger the antimicrobial activity. Organic germanium compounds are used by Horiuchi and Isamu for bacteriostasis and fungus inhibition tests, and the organic germanium can inhibit the growth and reproduction of fusarium, escherichia coli, staphylococcus aureus and bacillus subtilis. The experiment of the Sian Gwan with germanium, tin and lead compounds shows that the elements can inhibit the propagation of bacteria, the inhibition effect is in direct proportion to the atomic number of the elements, and the germanium has the strongest capacity of inhibiting the bacteria in the propagation period.
(3) Studies of removing free radicals and resisting oxidation, Zhang Paeonia, and the like show that a certain amount of germanium added into the low-selenium feed can obviously reduce the free radicals in the liver and the kidney of a rat. Harish tests on rats with low concentrations of lactic acid-germanium citrate found that glutathione peroxidase was significantly elevated in rat hepatocytes.
(4) The growth promoting germanium can enhance the metabolism function of the organism by increasing the content of iodine, triiodothyronine (T3) and tetraiodothyroxine (T4), thereby promoting the growth and development of animals and generating the weight increasing effect. The Yangxiangqiang and Tangmegaxin prove that the germanium has the functions of promoting the growth of broiler chickens and improving the dressing percentage and the feed conversion rate.
(5) The anticancer effect of Zongteng, Shimauchi, Suzuki and the like shows that the organic germanium can obviously prolong the survival time of the Ehrlich ascites type liver cancer mouse. Tangbo et al reported that germanium yeast has an inhibitory effect on liver cancer induced by aflatoxin B1.
Although the germanium compound has multiple biological functions of cancer resistance, oxidation resistance, bacteriostasis and the like, the biological toxicity of germanium must be noticed, the germanium compound has two types of inorganic germanium and organic germanium, the inorganic germanium has generally higher toxicity, and the application of the germanium in feed and food is also restricted; the organic germanium compound has lower chemical side effect and relatively high thermal stability, so the organic germanium has great application potential.
The organic germanium can be classified into synthetic organic germanium, natural organic germanium and biological organic germanium. The synthetic organic germanium is hydroxyethyl germanium sesquioxide, namely antiviral, anti-inflammatory and anticancer organic germanium such as Ge-132, spirogermanium, germanium furanose derivatives and the like, but calcium deficiency is easily caused by excessive administration; the natural organic germanium is extracted from natural plants, can be directly eaten, and has the defects of high cost and very limited yield; the biological organic germanium is obtained by enriching germanium compounds in organisms, such as yeast, bacteria, macrofungi and the like. Compared with natural organic germanium and synthetic organic germanium, the cost of synthesizing the biological organic germanium by utilizing microorganisms is lower, and the comprehensive benefit is higher.
The yeast is a mature microorganism, has the excellent performance of enriching trace elements, and is a strain which is most used for enriching elements at present. The candida utilis is a yeast which can grow by taking urea and nitric acid as nitrogen sources without adding any growth factor into a culture medium. It can use pentose and hexose, and can also use industrially cheap molasses, etc. as raw materials.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention mainly aims to provide a preparation method of germanium-rich yeast. The invention obtains the candida utilis with strong capacities of tolerating germanium, converting germanium and enriching germanium through domestication, and finally obtains the germanium-rich yeast with high biological activity and low toxicity by utilizing the capacities of the candida utilis of rich germanium and cheap raw materials and taking molasses and urea as fermentation raw materials. The method can convert inorganic germanium into organic germanium.
Another object of the present invention is a germanium-rich yeast obtained by the above production method.
It is still another object of the present invention to provide the use of the germanium-rich yeast.
The purpose of the invention is realized by the following technical scheme:
the invention uses cheap molasses and urea as fermentation substrates to prepare germanium-rich yeast, and the selected yeast is candida utilis. Obtaining a candida utilis strain with strong germanium tolerance, germanium conversion and germanium enrichment capacities through primary screening and secondary screening; and then obtaining a germanium-rich yeast product with high germanium content through an optimized fermentation process, wherein the germanium content is 837.1-1206.4 mg/kg, and the organic germanium content is 93.3-98.6%.
A preparation method of germanium-rich yeast comprises the following steps:
firstly, carrying out subculture and acclimatization on Candida utilis (Candida utilis) in a germanium-containing culture medium to obtain the acclimatized Candida utilis; then, the domesticated candida utilis is cultured in a yeast fermentation culture medium to obtain germanium-rich yeast.
The formula of the germanium-containing culture medium is preferably 50-150 g/L of molasses, 3-10 g/L of urea and 500-2000 mg/L of germanium dioxide;
the preferred subculture is subculture for 12-18 generations;
the domestication condition is preferably domestication at 25-35 ℃;
the formula of the yeast fermentation medium is preferably 50-80 g/L of molasses, 2-10 g/L of urea and 50-200 mg/L of germanium dioxide, and the pH value is 4.0-6.5; more preferably 50-80 g/L of molasses, 2-6 g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; more preferably 60g/L of molasses, 4g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; most preferably 60g/L of molasses, 4g/L of urea and 100mg/L of germanium dioxide, wherein the pH value is 4.0-6.5;
the culture conditions are preferably at 25 ℃ to 35 ℃.
A preparation method of germanium-rich yeast specifically comprises the following steps:
(1) subculturing candida utilis in a germanium-containing culture medium for acclimatization to obtain acclimatized candida utilis;
(2) performing amplification culture on the domesticated candida utilis obtained in the step (1) through slant strain culture, primary liquid seed culture, secondary liquid seed culture and fermentation tank seed culture;
(3) inoculating the culture solution obtained in the step (2) after the expanded culture into a yeast fermentation culture medium for fermentation, and drying the yeast fermentation liquor to obtain the germanium-rich yeast.
The formula of the germanium-containing culture medium in the step (1) is preferably 50-150 g/L of molasses, 3-10 g/L of urea and 500-2000 mg/L of germanium dioxide;
the preferred subculture is subculture for 12-18 generations;
the condition for acclimatization in the step (1) is preferably acclimatization at 25-35 ℃;
the culture medium for slant strain culture in the step (2) is preferably a potato sucrose culture medium, and the formula of the culture medium is 10-30 g/L of sucrose, 4-10 g/L of peptone, 5-10 g/L of yeast powder, 50-200 mg/L of germanium dioxide, and 18-20 g/L, pH of agar is 4.8-6.5; the potato sucrose liquid culture medium only needs no agar.
The culture condition of the slant strain culture in the step (2) is preferably culture at 25-35 ℃ for 45-50 h;
the culture medium for the primary liquid seed culture in the step (2) is preferably a potato sucrose liquid culture medium, and the formula of the culture medium is sucrose 20-40 g/L, peptone 4-10 g/L, yeast powder 5-10 g/L, germanium dioxide 50-200 mg/L, and agar 18-20 g/L, pH-4.8-6.5;
the first-stage liquid seed culture in the step (2) is preferably cultured for 54-66 h at 25-35 ℃;
performing primary liquid seed culture in the step (2), wherein 2-10 rings/L of hyphae obtained by slant strain culture are inoculated during culture;
the culture medium for the secondary liquid seed culture in the step (2) is preferably a potato sucrose liquid culture medium, and the potato sucrose liquid culture medium comprises 20-40 g/L of sucrose, 4-10 g/L of peptone, 5-10 g/L of yeast powder, 50-200 mg/L of germanium dioxide, and 18-20 g/L, pH of agar which is 4.8-6.5;
the secondary liquid seed culture in the step (2) is preferably cultured for 24-96 h at 25-35 ℃;
performing secondary liquid seed culture in the step (2), and inoculating 1% (V/V) of primary liquid seed culture during culture to obtain a primary seed culture solution;
the culture medium for seed culture of the fermentation tank in the step (2) is preferably a yeast fermentation culture medium, the formula of the yeast fermentation culture medium is preferably 50-80 g/L of molasses, 2-6 g/L of urea and 50-200 mg/L of germanium dioxide, and the pH value is 4.0-6.5; more preferably 50-80 g/L of molasses, 2-6 g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; more preferably 60g/L of molasses, 4g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; most preferably 60g/L of molasses, 4g/L of urea and 100mg/L of germanium dioxide, wherein the pH value is 4.0-6.5;
the culture condition of the seed culture of the fermentation tank in the step (2) is preferably culture at 25-35 ℃ for 24-96 h;
performing seed culture in the fermentation tank in the step (2), inoculating 5% (V/V) of secondary liquid seed culture during culture to obtain a secondary seed culture solution;
the formula of the yeast fermentation medium in the step (3) is preferably 50-80 g/L of molasses, 2-10 g/L of urea and 50-200 mg/L of germanium dioxide, and the pH value is 4.0-6.5; more preferably 50-80 g/L of molasses, 2-6 g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; more preferably 60g/L of molasses, 4g/L of urea and 100-150 mg/L of germanium dioxide, wherein the pH value is 4.0-6.5; most preferably 60g/L of molasses, 4g/L of urea and 100mg/L of germanium dioxide, wherein the pH value is 4.0-6.5;
the inoculation in the step (3) is preferably to inoculate the culture solution obtained in the step (2) after the expanded culture into a yeast fermentation culture medium according to the inoculation amount of 5 percent (V/V);
the fermentation condition in the step (3) is preferably fermentation at 25-35 ℃ for 24-96 h;
the drying in the step (3) is preferably spray drying, powder spraying drying, steam pressure roller drying or freeze drying; more preferably spray drying;
a germanium-rich yeast is prepared by the above preparation method. The germanium content in the germanium-rich yeast product is 837.1 mg/kg-1206.4 mg, wherein the content of organic germanium reaches 93.3% -98.6%.
Preferably, the germanium content in the germanium-rich yeast product is 1019.8 mg/kg-1206.4 mg, wherein the organic germanium content reaches 96.5% -98.2%.
The germanium-rich yeast is applied to the fields of food, feed and the like.
Compared with the prior art, the invention has the following advantages and effects:
(1) the invention utilizes yeast to convert inorganic germanium into organic germanium, and obtains the organic germanium with low biological toxicity.
(2) On the basis of a large number of experiments, the candida utilis strain with strong germanium tolerance, germanium conversion and germanium enrichment capacities is obtained, cheap molasses and urea can be used as fermentation substrates of the strain, and the production cost of organic germanium can be greatly reduced.
(3) On the basis of a large number of experiments, the method firstly cultures primary seeds of the candida utilis, then accesses a secondary seed solution containing germanium to perform expanded culture, and finally accesses a fermentation culture medium to prepare the germanium-rich microecological preparation, so that the microecological preparation is ensured to contain higher effective viable count of the germanium-rich yeast, and the tolerance strength and enrichment capacity of the yeast to the germanium are enhanced.
(4) The germanium-rich yeast product obtained by the invention has higher germanium content, the prior art is generally below 1000mg/kg, and the germanium-rich yeast product can reach 1206.4mg/kg under the most suitable condition.
(5) The method can convert inorganic germanium into organic germanium, expand the source of the organic germanium and promote the large-scale application of the germanium in the fields of food, feed and the like.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1: strain screening
Candida utilis (strain is purchased from China industrial microorganism strain preservation management center, and the strain number is CICC 1314), the strain is inoculated into a germanium-containing culture medium, subculture is respectively carried out on the strain under proper conditions (subculture is carried out for 12-18 generations) for domestication, and the enrichment condition of the strain on germanium is detected. Finally, obtaining a strain with strong germanium tolerance, germanium transformation and germanium enrichment capacity, namely the domesticated candida utilis.
The formula of the germanium-containing culture medium is as follows: 50-150 g/L of molasses, 3-10 g/L of urea and 500-2000 mg/L of germanium dioxide.
The culture conditions are as follows: the temperature is 25-35 ℃.
Example 2: preparation method of germanium-rich yeast
The production flow is as follows: slant culture → first liquid seed culture → second liquid seed culture → fermenter culture → separation → drying → packaging.
(1) Slant culture of strain: the domesticated candida utilis obtained in the example 1 is transferred into a potato sucrose culture medium (sucrose is 30g/L, peptone is 10g/L, yeast powder is 10g/L, germanium dioxide is 50mg/L, and agar is 20g/L, pH, the value is 4.8-6.5), the candida utilis is activated on a solid plate, the candida utilis is cultured at a constant temperature of 25-35 ℃ for 45 hours, and a single colony inclined surface is selected and stored for later use.
(2) First-stage liquid seed culture: 1-4 rings of yeast strains on the inclined plane are selected and inoculated in a triangular flask filled with 100-500 mL of potato sucrose liquid culture medium (30 g/L of sucrose, 10g/L of peptone, 10g/L of yeast powder, 50mg/L of germanium dioxide, and the pH value is 4.8-6.5), and the mixture is cultured at the constant temperature of 25-35 ℃ for 54 hours to obtain a first-level seed culture solution.
(3) Secondary liquid seed culture: inoculating the first-level seed culture solution into a Kaschin flask filled with 10-40L of potato sucrose liquid culture medium (30 g/L of sucrose, 10g/L of peptone, 10g/L of yeast powder, 50mg/L of germanium dioxide and 4.8-6.5 of pH value) according to the inoculation amount of 1% (V/V), and culturing at 25-35 ℃ for 24-96 h to obtain a second-level seed culture solution.
(4) Seed culture in a fermentation tank: inoculating the second-level seed culture solution into a culture medium containing 1-15 m of the second-level seed culture solution according to the inoculation amount of 5% (V/V)3Culturing in a seed fermentation tank of a yeast fermentation culture medium (molasses 80g/L, urea 6g/L, germanium dioxide 50mg/L, pH value of 4.0-6.5) at 26-35 ℃ for 24-96 h to obtain a fermentation tank seed culture solution.
(5) Culturing in a fermentation tank: inoculating seed culture solution of a fermentation tank to 15-180 m of seed culture solution according to the inoculation amount of 5% (V/V)3Yeast fermentation medium(molasses 80g/L, urea 6g/L, germanium dioxide 50mg/L, pH 4.0-6.5) in a fermentation tank, and performing fermentation culture at 25-35 ℃ for 24-96 h to obtain yeast fermentation liquor.
(6) Separation: separating with a laminated separator, and filtering with a vacuum drum or plate-frame filter press.
(7) And (3) drying: drying by spray drying method, and the dried product has activity; the powder spraying drying or the steam pressure roller drying is adopted, and the dried product has no activity and is powdery or flaky; or freeze drying.
(8) Packaging: various forms of packaging can be employed to produce the germanium-enriched yeast product. Wherein the content of germanium reaches 940.8mg/kg, and the content of organic germanium reaches 93.3%.
Example 3: preparation method of germanium-rich yeast
The production flow is as follows: slant culture → first liquid seed culture → second liquid seed culture → fermenter culture → separation → drying → packaging.
(1) Slant culture of strain: the domesticated candida utilis obtained in the example 1 is transferred into a potato sucrose culture medium (20 g/L of sucrose, 4g/L of peptone, 5g/L of yeast powder, 100mg/L of germanium dioxide and 20g/L of agar, the pH value is 4.8-6.5), activated on a solid plate, cultured at the constant temperature of 28 ℃ for 48 hours, and a single colony inclined surface is selected for storage and standby.
(2) First-stage liquid seed culture: 1-4 rings of yeast strains on the inclined plane are selected and inoculated in a triangular flask filled with 100-500 mL of potato sucrose liquid culture medium (20 g/L of sucrose, 4g/L of peptone, 5g/L of yeast powder, 100mg/L of germanium dioxide and pH value of 4.8-6.5), and the mixture is cultured for 60 hours at constant temperature of 28 ℃ to obtain a primary seed culture solution.
(3) Secondary liquid seed culture: inoculating the first-level seed culture solution into a Kaschin flask filled with 10-40L of potato sucrose liquid culture medium (20 g/L of sucrose, 4g/L of peptone, 5g/L of yeast powder, 100mg/L of germanium dioxide and 4.8-6.5 of pH value) according to the inoculation amount of 1% (V/V), and culturing at 25-35 ℃ for 24-96 h to obtain a second-level seed culture solution.
(4) Seed culture in a fermentation tank: the second class seedInoculating the seed culture solution to a seed container with a volume of 1-15 m according to an inoculation amount of 5% (V/V)3Culturing in a seed fermentation tank with a yeast fermentation culture medium (60 g/L of molasses, 4g/L of urea, 100mg/L of germanium dioxide and pH value of 4.0-6.5) at 25-35 ℃ for 24-96 h to obtain a seed culture solution of the fermentation tank.
(5) Culturing in a fermentation tank: inoculating seed culture solution of a fermentation tank to 15-180 m of seed culture solution according to the inoculation amount of 5% (V/V)3Fermenting and culturing for 24-96 h at 25-35 ℃ in a fermentation tank of a yeast fermentation medium (60 g/L of molasses, 4g/L of urea, 100mg/L of germanium dioxide and pH value of 4.0-6.5) to obtain yeast fermentation liquor.
(6) Separation: separating with a laminated separator, and filtering with a vacuum drum or plate-frame filter press.
(7) And (3) drying: drying by spray drying method, and the dried product has activity; the powder spraying drying or the steam pressure roller drying is adopted, and the dried product has no activity and is powdery or flaky; or freeze drying.
(8) Packaging: various forms of packaging can be employed to produce the germanium-enriched yeast product. Wherein the content of germanium reaches 1206.4mg/kg, and the content of organic germanium reaches 96.5%.
Example 4: preparation method of germanium-rich yeast
The production flow is as follows: slant culture → first liquid seed culture → second liquid seed culture → fermenter culture → separation → drying → packaging.
(1) Slant culture of strain: the domesticated candida utilis obtained in the example 1 is transferred into a potato sucrose culture medium (10 g/L of sucrose, 6g/L of peptone, 8g/L of yeast powder, 150mg/L of germanium dioxide, 20g/L of agar, and the pH value is 4.8-6.5), activated on a solid plate, cultured at the constant temperature of 25-35 ℃ for 50 hours, and a single colony inclined surface is selected for storage and standby.
(2) First-stage liquid seed culture: and (3) selecting 1-4 rings of yeast strains on the inclined plane, inoculating the yeast strains into a triangular flask filled with 100-500 mL of potato sucrose liquid culture medium (20 g/L of sucrose, 6g/L of peptone, 8g/L of yeast powder, 150mg/L of germanium dioxide and 4.8-6.5 of pH value), and culturing at the constant temperature of 25-35 ℃ for 60 hours to obtain a primary seed culture solution.
(3) Secondary liquid seed culture: inoculating the first-level seed culture solution into a Kaschin flask filled with 10-40L of potato sucrose liquid culture medium (20 g/L of sucrose, 6g/L of peptone, 8g/L of yeast powder, 150mg/L of germanium dioxide and 4.8-6.5 of pH value) according to the inoculation amount of 1% (V/V), and culturing at 25-35 ℃ for 24-96 h to obtain a second-level seed culture solution.
(4) Seed culture in a fermentation tank: inoculating the second-level seed culture solution into a culture medium containing 1-15 m of the second-level seed culture solution according to the inoculation amount of 5% (V/V)3Culturing in a seed fermentation tank of a yeast fermentation culture medium (50 g/L of molasses, 2g/L of urea, 150mg/L of germanium dioxide and pH value of 4.0-6.5) at 25-35 ℃ for 24-96 h to obtain a seed culture solution of the fermentation tank.
(5) Culturing in a fermentation tank: inoculating seed culture solution of a fermentation tank to 15-180 m of seed culture solution according to the inoculation amount of 5% (V/V)3Fermenting and culturing for 24-96 h at 25-35 ℃ in a fermentation tank of a yeast fermentation medium (50 g/L of molasses, 2g/L of urea, 150mg/L of germanium dioxide and pH value of 4.0-6.5) to obtain yeast fermentation liquor.
(6) Separation: separating with a laminated separator, and filtering with a vacuum drum or plate-frame filter press.
(7) And (3) drying: drying by spray drying method, and the dried product has activity; the powder spraying drying or the steam pressure roller drying is adopted, and the dried product has no activity and is powdery or flaky; or freeze drying.
8) Packaging: various forms of packaging can be employed to produce the germanium-enriched yeast product. Wherein the content of germanium reaches 1019.8mg/kg, and the content of organic germanium reaches 98.2%.
Example 5: preparation method of germanium-rich yeast
The production flow is as follows: slant culture → first liquid seed culture → second liquid seed culture → fermenter culture → separation → drying → packaging.
(1) Slant culture of strain: the domesticated candida utilis obtained in the example 1 is transferred into a potato sucrose culture medium (25 g/L of sucrose, 4g/L of peptone, 6g/L of yeast powder, 200mg/L of germanium dioxide and 20g/L, pH of agar, the value is 4.8-6.5), activated on a solid plate, cultured at the constant temperature of 25-35 ℃ for 48 hours, and a single colony inclined surface is selected for storage.
(2) First-stage liquid seed culture: and (3) selecting 1-4 rings of yeast strains on the inclined plane, inoculating the yeast strains into a triangular flask filled with 100-500 mL of potato sucrose liquid culture medium (40 g/L of sucrose, 4g/L of peptone, 6g/L of yeast powder, 200mg/L of germanium dioxide, and the pH value of the culture medium is 4.8-6.5), and culturing at the constant temperature of 25-35 ℃ for 66 hours to obtain a primary seed culture solution.
(3) Secondary liquid seed culture: inoculating the first-level seed culture solution into a Kaschin flask filled with 10-40L of potato sucrose liquid culture medium (sucrose 40g/L, peptone 4g/L, yeast powder 6g/L, germanium dioxide 200mg/L and pH value of 4.8-6.5) according to the inoculation amount of 1% (V/V), and culturing at 25-35 ℃ for 24-96 h to obtain a second-level seed culture solution.
(4) Seed culture in a fermentation tank: inoculating the second-level seed culture solution into a culture medium containing 1-15 m of the second-level seed culture solution according to the inoculation amount of 5% (V/V)3Culturing in a seed fermentation tank of a yeast fermentation culture medium (70 g/L of molasses, 4g/L of urea, 200mg/L of germanium dioxide and pH value of 4.0-6.5) at 25-35 ℃ for 24-96 h to obtain a seed culture solution of the fermentation tank.
(5) Culturing in a fermentation tank: inoculating seed culture solution of a fermentation tank to 15-180 m of seed culture solution according to the inoculation amount of 5% (V/V)3Fermenting and culturing for 24-96 h at 25-35 ℃ in a fermentation tank of a yeast fermentation medium (molasses 80g/L, urea 10g/L, germanium dioxide 200mg/L and pH value of 4.0-6.5) to obtain yeast fermentation liquor.
(6) Separation: separating with a laminated separator, and filtering with a vacuum drum or plate-frame filter press.
(7) And (3) drying: drying by spray drying method, and the dried product has activity; the powder spraying drying or the steam pressure roller drying is adopted, and the dried product has no activity and is powdery or flaky; or freeze drying.
(8) Packaging: various forms of packaging can be employed to produce the germanium-enriched yeast product. Wherein the content of germanium reaches 837.1mg/kg, and the content of organic germanium reaches 98.6%.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (5)
1. A preparation method of germanium-rich yeast is characterized by comprising the following steps:
first, Candida utilis (A), (B) andCandida utilis) Subculturing and domesticating the CICC 1314 in a germanium-containing culture medium to obtain domesticated candida utilis; then, culturing the domesticated candida utilis in a yeast fermentation culture medium to obtain germanium-rich yeast;
the formula of the germanium-containing culture medium comprises 50-150 g/L of molasses, 3-10 g/L of urea and 500-2000 mg/L of germanium dioxide;
the formula of the yeast fermentation medium comprises 50-80 g/L of molasses, 2-10 g/L of urea and 50-200 mg/L of germanium dioxide, and the pH is = 4.0-6.5;
the method comprises the following specific steps:
(1) subculturing candida utilis in a germanium-containing culture medium for acclimatization to obtain acclimatized candida utilis;
(2) performing amplification culture on the domesticated candida utilis obtained in the step (1) through slant strain culture, primary liquid seed culture, secondary liquid seed culture and fermentation tank seed culture; the culture medium for seed culture in the fermentation tank is a yeast fermentation culture medium;
the culture medium for slant strain culture is a potato sucrose culture medium, and the formula of the culture medium is 10-30 g/L of sucrose, 4-10 g/L of peptone, 5-10 g/L of yeast powder, 50-200 mg/L of germanium dioxide and 18-20 g/L, pH = 4.8-6.5 of agar; the potato sucrose liquid culture medium only needs not to be added with agar;
the first-stage liquid seed culture medium is a potato sucrose liquid culture medium, and the formula of the first-stage liquid seed culture medium is 20-40 g/L of sucrose, 4-10 g/L of peptone, 5-10 g/L of yeast powder, 50-200 mg/L of germanium dioxide and 18-20 g/L, pH = 4.8-6.5 of agar;
the culture medium for the secondary liquid seed culture is a potato sucrose liquid culture medium, and the formula of the culture medium is 20-40 g/L of sucrose, 4-10 g/L of peptone, 5-10 g/L of yeast powder, 50-200 mg/L of germanium dioxide and 18-20 g/L, pH = 4.8-6.5 of agar;
the culture condition of the slant strain culture is 25-35 ℃ for 45-50 h;
the first-stage liquid seed culture is carried out under the culture condition of 25-35 ℃ for 54-66 h;
the culture condition of the secondary liquid seed culture is that the secondary liquid seed culture is cultured for 24-96 h at 25-35 ℃;
the culture condition of the seed culture of the fermentation tank is that the seed culture is carried out for 24-96 h at 25-35 ℃;
(3) inoculating the culture solution obtained in the step (2) after the expanded culture into a yeast fermentation culture medium for fermentation, and drying the yeast fermentation liquid to obtain germanium-rich yeast; the fermentation condition is fermentation at 25-35 ℃ for 24-96 h.
2. The method for producing a germanium-rich yeast according to claim 1, wherein:
the culture medium for seed culture of the fermentation tank in the step (2) is a yeast fermentation culture medium, and the formula of the yeast fermentation culture medium comprises 50-80 g/L of molasses, 2-6 g/L of urea and 50-200 mg/L of germanium dioxide, and the pH = 4.0-6.5.
3. The method for producing a germanium-rich yeast according to claim 1, wherein:
the drying in the step (3) is spray drying, powder spraying drying, steam pressure roller drying or freeze drying.
4. A germanium-rich yeast characterized by: prepared by the preparation method of any one of claims 1 to 3.
5. Use of the germanium-rich yeast of claim 4 for the preparation of a food or feed product.
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|---|---|---|---|---|
| JPS53130483A (en) * | 1977-04-20 | 1978-11-14 | Gerumatsukusu Kk | Production of yeasts containing germanium |
| US5792646A (en) * | 1993-11-29 | 1998-08-11 | Daijy Corporation | Process for preparing S. cerevisiae containing organically bound germanium |
| KR20050108103A (en) * | 2004-05-11 | 2005-11-16 | 주식회사 엔지켐 | Method for producing yeast including organically bound germanium and selenium |
| KR20080070894A (en) * | 2007-01-29 | 2008-08-01 | 주식회사 한국수리미 | Production method of functional feed additives for fish culturing and feed additives from this method |
| CN105602861A (en) * | 2015-10-20 | 2016-05-25 | 珲春市龙裕农业发展集团有限公司 | Germanium-rich yeast additive adopting rice bran as medium, and preparation method thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS53130483A (en) * | 1977-04-20 | 1978-11-14 | Gerumatsukusu Kk | Production of yeasts containing germanium |
| US5792646A (en) * | 1993-11-29 | 1998-08-11 | Daijy Corporation | Process for preparing S. cerevisiae containing organically bound germanium |
| KR20050108103A (en) * | 2004-05-11 | 2005-11-16 | 주식회사 엔지켐 | Method for producing yeast including organically bound germanium and selenium |
| KR20080070894A (en) * | 2007-01-29 | 2008-08-01 | 주식회사 한국수리미 | Production method of functional feed additives for fish culturing and feed additives from this method |
| CN105602861A (en) * | 2015-10-20 | 2016-05-25 | 珲春市龙裕农业发展集团有限公司 | Germanium-rich yeast additive adopting rice bran as medium, and preparation method thereof |
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